Department of Neurology, Children's Hospital of Hebei Province, Shijiazhuang, Hebei, China.
Kaohsiung J Med Sci. 2019 Sep;35(9):527-534. doi: 10.1002/kjm2.12102. Epub 2019 Aug 2.
Epilepsy is one of the most common neurological disorders in humans. Recently, long noncoding RNAs (lncRNAs) have been reported to be important players in neurological diseases. Herein, this study aimed to examine the effect of lncRNA GAS5 on the occurrence of epilepsy in rat and cell models of epileptic seizure. The expression of lncRNA GAS5 was measured in the established rat and cell models. The binding sites between lncRNA GAS5 and miR-135a-5p, as well as those between miR-135a-5p and 3' untranslated region of KCNQ3 were predicted by miRDB and Targetscan, separately, followed by verification using dual-luciferase reporter gene assay. The expression of miR-135a-5p was measured in response to the overexpression of lncRNA GAS5. The mRNA and protein levels of KCNQ3 were examined in response to overexpression of miR-135a-5p. Next, the latency of epilepsy and frequency of epileptic seizures were assessed in rats injected with Lv-shGAS5 and Lv-miR-135a-5p in epileptic seizure model. In the rat and cell models, lncRNA GAS5 was highly expressed when epileptic seizure was induced. The expression of miR-135a-5p was decreased by overexpression of lncRNA GAS5. Meanwhile, the mRNA and protein levels of KCNQ3 were decreased in response to knockdown of miR-135a-5p. After the treatment of Lv-shGAS5 and Lv-miR-135a-5p, the average latent period of epilepsy was prolonged and the frequency of seizures was decreased. The key findings of the present study provide evidence emphasizing that lncRNA GAS5 functions as a competitive endogenous RNA of miR-135a-5p to increase expression of KCNQ3, and lncRNA GAS5 silencing inhibited the occurrence and progression of epilepsy.
癫痫是人类最常见的神经障碍之一。最近，长链非编码 RNA（lncRNA）已被报道在神经疾病中发挥重要作用。本研究旨在探讨 lncRNA GAS5 对大鼠和癫痫发作细胞模型中癫痫发生的影响。建立大鼠和细胞模型，检测 lncRNA GAS5 的表达。分别通过 miRDB 和 Targetscan 预测 lncRNA GAS5 与 miR-135a-5p 以及 miR-135a-5p 与 KCNQ3 3'非翻译区之间的结合位点，然后通过双荧光素酶报告基因检测进行验证。检测 lncRNA GAS5 过表达时 miR-135a-5p 的表达情况。检测过表达 miR-135a-5p 时 KCNQ3 的 mRNA 和蛋白水平。接下来，在癫痫发作模型中注射 Lv-shGAS5 和 Lv-miR-135a-5p 评估大鼠的癫痫潜伏期和癫痫发作频率。在大鼠和细胞模型中，诱导癫痫发作时 lncRNA GAS5 表达升高。lncRNA GAS5 过表达可降低 miR-135a-5p 的表达。同时，敲低 miR-135a-5p 可降低 KCNQ3 的 mRNA 和蛋白水平。经 Lv-shGAS5 和 Lv-miR-135a-5p 治疗后，癫痫潜伏期延长，发作频率降低。本研究的主要发现为长链非编码 RNA GAS5 作为 miR-135a-5p 的竞争性内源性 RNA，增加 KCNQ3 的表达，沉默 lncRNA GAS5 可抑制癫痫的发生和发展提供了证据。
